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Method for evaluating the degree of maturity of corneocytes

Inactive Publication Date: 2008-06-12
HIRAO TETABUJI +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0008]The present inventors have been repeating researches paying attentions to a horny layer barrier function, particularly the relation of CE to intercellular lipids which is considered to mainly constitute barrier function to prevent water evaporation from the body and invasion of foreign substances from the outside and the properties of CE. It has been confirmed that evaluation of a corneocyte suitably prepared causes acquisition of hydrophobicity of CE and a change in disappearance of ant

Problems solved by technology

However, what a remaining nucleus means is not necessarily distinct, and it has not yet scientifically been explained particularly in relation to a barrier function.

Method used

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  • Method for evaluating the degree of maturity of corneocytes
  • Method for evaluating the degree of maturity of corneocytes
  • Method for evaluating the degree of maturity of corneocytes

Examples

Experimental program
Comparison scheme
Effect test

example 1

Evaluation of a Horny Layer Sample Obtained Directly from the Skin

[0037]A cellophane adhesive tape was adhered on a face (cheek) and an upper arm of an examinee having skin troubles such as rough skin and immediately peeled off. A Tris-hydrochloric acid buffer solution containing dithiothreitol and sodium dodecylsulfate (SDS) was added to the horny layer adhered to the tape, and the mixture was heated at 100° C. for 10 minutes. Insoluble substances were obtained by centrifugation of 4000×g for 10 minutes. Further, adding and heating of the effluent were repeated three times to thoroughly remove the soluble components. The insoluble substances thus obtained was CE.

[0038]Respective CE's in horny layer samples originating in the upper arm inside (non-processed group) of the examinee which was prepared by the method described above, an SDS-induced rough skin group and a tape stripping-induced rough skin group each were dropped on a slide glass, dried by air and then fixed in cold aceton...

example 2

Evaluation of Cultured Epidermal Keratinocytes

[0042]A human epidermal keratinocytes were cultured according to a Rheinwald & Green method (Cell: 6: 331-334, 1975). It was cultured in a growth medium (DMEM-Ham's sF 12 (3:1) containing 10% fetal calf serum, hydrocortisone 0.5 μg / ml, insulin 5 μg / ml, cholera toxin 10−10 M, epidermal growth factor 10 ng / ml and adenine 1.8×10−4 M) and reached confluent, and then the medium was substituted with a culture medium (MEM containing 0.1% bovine serum albumin) containing oleic acid or linoleic acid which is known to urge the differentiation of keratinocite to accelerate the formation of a barrier (Hanley et al., Journal of Clinical Investigation 100: 705-712, 1997 and Komuves et al., Journal of Investigative Dermatology 111: 429-433, 1998). Further, culture was continued for one week. After finishing the culture, a Tris-hydrochloric acid buffer solution containing dithiothreitol and sodium dodecylsulfate (SDS) was added thereto, and the mixture ...

example 3

Detection of Nuclear Component in Corneocyte

[0043]The corneocyte was dispersed from the horny layer sample according to the method of Takahashi et al. (described above). That is, a cellophane adhesive tape was adhered on the skin of a part to be examined and immediately peeled off. A sodium dodecylsulfate (SDS)-dodecyldimethylamine oxide (C12DMAO) mixed solution was added to the horny layer adhered to the tape and heated at 50° C. for 24 hours. The dispersed corneocytes were obtained by centrifugation of 4000×g for 10 minutes. Further, they were washed three times repeatedly with the SDS-C12DMAO mixed solution to thoroughly remove the soluble components to thereby prepare the corneocytes.

[0044]The corneocytes prepared by the method described above were dropped on a slide glass, dried by air and then fixed in cold acetone. They were hydrated in a Dulbecco's phosphate buffered saline and then reacted with a mouse anti-involucrin antibody (NOVOCASTRA Co., Ltd.) used as a primary antibo...

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Abstract

Disclosed is an evaluating method for the properties of a corneocyte in a horny layer sample originating in the skin. In the above method, detected and evaluated are selective staining of a cornified envelope in a corneocyte, selective staining of a nuclear component and the antigenicity.

Description

TECHNICAL FIELD[0001]The present invention relates to a method for evaluating the degree of maturity of a corneocyte constituting a horny layer of the skin. More specifically, the present invention relates to a method for evaluating a skin quality, which is useful in the field of a making-up method or a dermatology and to a kit used for the above evaluating method.BACKGROUND ART[0002]It is important in taking precise skin care for maintaining the healthier skin to accurately grasp the skin quality (or the condition of the skin). Accordingly, the skin quality of users of cosmetics has so far been evaluated through, for example, an inquiry made by a beauty technician in carrying out skin care by cosmetics. Further, the condition or function of the skin has so far been evaluated by parameters observed or measured by means of various measuring equipments for the purpose of objectively evaluating the skin quality.[0003]The representatives of such parameters include a skin surface morphol...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N33/569
CPCG01N1/30G01N33/56966G01N33/5091G01N33/5005
Inventor HIRAO, TETSUJIDENDA, MITSUHIROTAKAHASHI, MOTOJITERUI, TADASHITAGAMI, HACHIRO
Owner HIRAO TETABUJI
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